Rxivist logo

Improving oligo-conjugated antibody signal in multimodal single-cell analysis

By Terkild B Buus, Alberto Herrera, Ellie Ivanova, Eleni P. Mimitou, Anthony Cheng, Ramin Sedaghat Herati, Thales Papagiannakopoulos, Peter Smibert, Niels Odum, Sergei B Koralov

Posted 16 Jun 2020
bioRxiv DOI: 10.1101/2020.06.15.153080

Simultaneous measurement of surface proteins and gene expression within single cells using oligo-conjugated antibodies offers high resolution snapshots of complex cell populations. Signal from oligo-conjugated antibodies is quantified by high-throughput sequencing and is highly scalable and sensitive. In this study, we investigated the response of oligo-conjugated antibodies towards four variables: Concentration, staining volume, cell number at staining, and tissue. We find that staining with recommended antibody concentrations cause unnecessarily high background and that concentrations can be drastically reduced without loss of biological information. Reducing volume only affects antibodies targeting abundant epitopes used at low concentrations and is counteracted by reducing cell numbers. Adjusting concentrations increases signal, lowers background and reduces costs. Background signal can account for a major fraction of the total sequencing and is primarily derived from antibodies used at high concentrations. Together, this study provides new insight into the titration response and background of oligo-conjugated antibodies and offers concrete guidelines on how such panels can be improved.

Download data

  • Downloaded 3,194 times
  • Download rankings, all-time:
    • Site-wide: 5,971
    • In genomics: 570
  • Year to date:
    • Site-wide: 25,889
  • Since beginning of last month:
    • Site-wide: 12,558

Altmetric data

Downloads over time

Distribution of downloads per paper, site-wide